Novel Tetragenococcus halophilus with high yield of umami peptide and application of Tetragenococcus halophilus

A halophilic tetrad, umami peptide technology, applied in the direction of bacteria, microorganism-based methods, food ingredients as taste improvers, etc., can solve the problem of high cost, low yield of production strains, and inability to meet the needs of large-scale industrial production, etc. problems, to achieve the effect of short production cycle, high output and improved safety

Active Publication Date: 2022-02-01
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few studies on the fermentation of umami peptides through various metabolic pathways of microorganisms. The production strains used have low

Method used

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  • Novel Tetragenococcus halophilus with high yield of umami peptide and application of Tetragenococcus halophilus
  • Novel Tetragenococcus halophilus with high yield of umami peptide and application of Tetragenococcus halophilus
  • Novel Tetragenococcus halophilus with high yield of umami peptide and application of Tetragenococcus halophilus

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Experimental program
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Effect test

Embodiment 1

[0030] The isolation and screening of embodiment 1 bacterial strain

[0031] 1. Screening samples

[0032] Soybean paste collected from Shangpingzhou Village, Xiaotun Town, Liaoyang City, which has been naturally fermented for half a year.

[0033] 2. Preliminary screening of tetradococci

[0034] Take 1g of soybean paste sample, add 9mL of 0.9% sterile normal saline, mix well and soak for 15min; then take 1mL of sample solution and inoculate it in the enrichment medium, culture it at 37℃ for 1-2d, and spread it on a flat plate Cultured on solid separation medium for 5-6d.

[0035] Pick a single colony that is small, has obvious calcium-dissolving circle, is milky white and opaque, and has a shape similar to that of a lactic acid bacteria colony, and performs Gram staining. Microscopic observation results showed that there were 12 bacterial strains in the form of pairs or quadruple globules, which were named LY9-1, LY9-2, ..., LY9-12 respectively.

[0036] Purify and cultu...

Embodiment 2

[0053] Identification of embodiment 2LY9-1 bacterial strain

[0054] 1. Identification of colony morphology

[0055] The bacterium colony of LY9-1 bacterial strain described in the present invention is less, protrudes, and is milky white, opaque, and the surface is smooth and glossy, and the edge is flat; No motility, does not produce spores; Gram staining is positive, and the cell is spherical, quadruple or in pairs. Colony morphology and cell morphology figure 1 and figure 2 shown.

[0056] 2. Molecular biological identification

[0057] (1) Bacterial DNA extraction:

[0058] Pipette 100 μL of the LY9-1 bacterial suspension into the sterilized MRS liquid medium, and culture it in a shaking incubator at 37°C for 24 hours. The genome of the strain was extracted according to the operating steps of the bacterial genome kit (Solarbio D1600).

[0059] (2) PCR amplification:

[0060] PCR upstream primer 27F and downstream primer 1492R were designed and synthesized by Shang...

Embodiment 3

[0073] Example 3 Determination of Enzyme Production Ability of Tetradynococcus halophilus SNTH-1

[0074] 1. Determination of protease activity:

[0075] After the halophilic Tetradendococcus SNTH-1 strain was activated, it was inoculated in the MRS liquid medium and cultured in a constant temperature incubator at 37°C for 24 hours.

[0076] Prepare protease screening medium (10g casein, 3g yeast powder, 5g Na 2 HPO 4 12H 2O, 50g NaCl, 0.05g bromothymol blue, 20g agar, 1L distilled water, adjust the pH value to 7), and adopt the Oxford cup method to measure the protease-producing ability of the bacterial strain. Put the sterilized Oxford cup on the protease screening medium, absorb 10 μ of the halophilic Tetradendococcus SNTH-1 bacteria solution into the Oxford cup, and cultivate it in a constant temperature incubator at 37°C for 24 hours, and observe whether there is a transparent circle.

[0077] The casein hydrolysis circle diameter (D) and the colony diameter (d) were ...

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Abstract

The invention relates to the technical field of functional microorganism screening and application, and particularly provides a strain of novel Tetragenococcus halophilus and application of the novel Tetragenococcus halophilus. The Tetragenococcus halophilus is screened from traditional naturally-fermented soybean paste in Northeast, and is preserved in China General Microbiological Culture Collection Center (CGMCC) on August 20, 2021, wherein the preservation address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No. 23165. The strain can greatly improve the yield of umami peptide, is beneficial to reducing the production cost of the umami peptide, and promotes the wide application of the umami peptide in the food field.

Description

technical field [0001] The invention relates to the technical field of functional microorganism screening and application, in particular to a novel high-yield umami peptide halophilic tetrad and its application. Background technique [0002] As a new type of umami substance proposed in recent years, umami peptide has good umami effect, processing characteristics, and nutritional value. widespread attention. Umami peptides are small molecular peptides with umami properties extracted from food or synthesized by amino acids, which can supplement and enhance the overall taste of food, making it more harmonious, soft and rich, and its molecular mass is usually 150 to 3000Da. The taste of umami peptides mainly comes from the intermediate products of protein synthesis and decomposition process. Compared with other peptides, umami peptide molecules usually contain glutamic acid residues or aspartic acid residues, and these molecules also interact with umami receptors on human tast...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23L27/24A23L29/00C12R1/01
CPCC12N1/20A23L27/24A23L29/065A23V2002/00A23V2250/55A23V2200/16
Inventor 乌日娜武俊瑞曹恺欣安飞宇赵越
Owner SHENYANG AGRI UNIV
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